Wang S Y, Mak K L, Chen L Y, Chou M P, Ho C K
Department of Medical Research, Veterans General Hospital-Taipei, Taiwan, Republic of China.
Immunology. 1992 Oct;77(2):298-303.
Human blood mononuclear cells from normal adults were collected after density-cut centrifugation and monocytes were then isolated by removal of lymphocytes using the techniques of E-rosetting and cell adhesion. The purified monocytes were further analysed by velocity sedimentation, and two distinct subpopulations with different cell sizes were obtained. The larger monocytes were 17.0 +/- 1.8 microns in diameter with a mean sedimentation rate (SR) of 7.0 +/- 0.6 mm/hr, while the smaller monocytes were 9.5 +/- 0.8 microns in size and 4.1 +/- 0.2 mm/hr in SR. The population ratio of larger:smaller cells was approximately 2:1 (66 +/- 2.8%:34 +/- 1.6%). Both cell populations exhibited a high positive rate (> 98%) in both the non-specific esterase and the peroxidase stain. However, the larger cells had much higher phagocytic activity than the smaller ones. Furthermore, the expression of monocyte-associated antigens was also different between these two subpopulations. Thus, while most of the larger monocytes (98%) could be recognized by monoclonal antibodies MY7 and OKM1, only some (35 and 61%, respectively) of the smaller monocytes could react with those antibodies. In addition, the larger monocytes secreted a significant amount of monokines including interleukin-1 beta (IL-1 beta), tumour necrosis factor-alpha (TNF-alpha) and prostaglandin E2 (PGE2) and their production increased in proportion to the level of stimulation by bacterial lipopolysaccharide (LPS), whereas the production of monokines by the smaller monocytes remained at low levels and did not respond to LPS stimulation. These results reveal the existence of phenotypic and functional heterogeneity in human blood monocytes.
通过密度梯度离心收集正常成年人的人血单核细胞,然后采用E花环形成和细胞黏附技术去除淋巴细胞以分离单核细胞。通过速度沉降对纯化的单核细胞进行进一步分析,获得了两个具有不同细胞大小的不同亚群。较大的单核细胞直径为17.0±1.8微米,平均沉降速率(SR)为7.0±0.6毫米/小时,而较小的单核细胞大小为9.5±0.8微米,SR为4.1±0.2毫米/小时。较大细胞与较小细胞的群体比例约为2:1(66±2.8%:34±1.6%)。两个细胞群体在非特异性酯酶和过氧化物酶染色中均表现出高阳性率(>98%)。然而,较大的细胞比较小的细胞具有更高的吞噬活性。此外,这两个亚群之间单核细胞相关抗原的表达也有所不同。因此,虽然大多数较大的单核细胞(98%)可被单克隆抗体MY7和OKM1识别,但只有一些(分别为35%和61%)较小的单核细胞能与这些抗体反应。此外,较大的单核细胞分泌大量的单核因子,包括白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和前列腺素E2(PGE2),它们的产生与细菌脂多糖(LPS)刺激水平成比例增加,而较小的单核细胞产生的单核因子水平较低且对LPS刺激无反应。这些结果揭示了人血单核细胞中存在表型和功能异质性。